title: Functional characterization of the trypanosome translational repressor SCD6.
abstract: The storage of translationally inactive mRNAs in cytosolic granules enables cells to react flexibly to environmental changes. In eukaryotes, Scd6 (suppressor of clathrin deficiency 6)/Rap55 (RNA-associated protein 55), a member of the LSm14 (like-Sm14) family, is an important factor in the formation and activity of P-bodies, where mRNA decay factors accumulate, in stress granules that store mRNAs under adverse conditions and in granules that store developmentally regulated mRNAs. SCD6 from Trypanosoma brucei (TbSCD6) shares the same domain architecture as orthologous proteins in other organisms and is also present in cytosolic granules (equivalent to P-bodies). We show that TbSCD6 is a general repressor of translation and that its depletion by RNAi results in a global increase in protein synthesis. With few exceptions, the steady-state levels of proteins are unchanged. TbSCD6 is not required for the formation of starvation-induced granules in trypanosomes, and unlike Scd6 from yeast, Plasmodium and all multicellular organisms analysed to date, it does not form a complex with the helicase Dhh1 (DExD/H-box helicase 1). In common with Xenopus laevis RAP55, TbSCD6 co-purifies with two arginine methyltransferases; moreover, TbSCD6 itself is methylated on three arginine residues. Finally, a detailed analysis identified roles for the Lsm and N-rich domains in both protein localization and translational repression.
authors.affiliation: *Institute of Cell Biology, University of Bern, Bern, Switzerland.
authors.forename: Marina
authors.initials: M
authors.lastname: Cristodero
authors.forename: Bernd
authors.initials: B
authors.lastname: Schimanski
authors.forename: Manfred
authors.initials: M
authors.lastname: Heller
authors.forename: Isabel
authors.initials: I
authors.lastname: Roditi
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D007553:Isotope Labeling / Q000379:methods; D030781:Organisms, Genetically Modified; D010641:Phenotype; D011485:Protein Binding; D014176:Protein Biosynthesis / Q000187:drug effects / Q000235:genetics*; D020543:Proteome / Q000032:analysis; D034741:RNA, Small Interfering / Q000494:pharmacology; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D012261:Ribonucleoproteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D014346:Trypanosoma brucei brucei / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D020543:Proteome; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D012261:Ribonucleoproteins